Project Summary/Abstract (Description) Consistent with the mission of the funding announcement to use engineering principles to solve problems in biomedical science, this research proposes to develop new tools to monitor hepatocellular carcinoma (HCC) based on changes in hepatic fructose metabolism. HCC is a malignancy of the liver and the prognosis of the disease does not solely depend on tumor stage but also on liver function. The prevalence of metabolic syndrome has also seen non-alcoholic fatty liver disease (NAFLD) as the leading cause of HCC in the US. While there is an increase incidence of this HCC, there is an unmet need for tolls that enable early detection and staging of HCC. This study proposes to develop new biomarkers to track cancer progression based on the loss of fructose metabolism in HCC. We hypothesize that fructose metabolism is lost in the development of liver cancer and can be used track HCC. This is based on published literature and our recent experiments showing proteins involved in hepatic fructose metabolism including ketohexokinase (KHK) are down regulated in HCC. These observations form the foundation of our hypothesis that fructose metabolism is the preserve of the normal liver and is down regulated in liver cancer. Therefore, development of a blood-based test to measure fructose levels in the blood will inform on hepatic function. A complementary metabolic imaging biomarker based on hyperpolarized magnetic resonance imaging (HP-MRI) will provide real-time, spatial information. HP-MRI is a new imaging technology that improves detection of 13C-labeled molecules, enabling non-invasive quantification of metabolic flux and is currently undergoing clinical trials. In Aim 1, we will measure blood and tissue levels of fructose after an intravenous fructose injection in a mouse model of HCC in a background of NAFLD. We will use mass spectrometry, in a new collaboration with the School of Pharmacy. We will then investigate the mechanism of action of this phenomenon by focusing on the transcription factor, hepatocyte nuclear factor 4 alpha (HNF4A). In Aim 2, we will optimize polarization of [2-13C] fructose, to enable detection of KHK activity in normal and HCC mice. This project proposes to develop innovative and quantitative methods to track HCC based on a novel biological observation of altered liver metabolism in HCC. Investigators from diverse backgrounds such as biochemistry, metabolomics and MR engineering have been assembled to ensure success. The completion of this project will deliver complementary, translatable technologies for monitoring HCC.